Graves病患者外周血免疫调节细胞及其亚群比例异常

目的观察Graves病(GD)患者外周血各种免疫调节细胞及其亚群的变化。方法采用流式细胞术(FCM)分别检测32例GD患者及30例健康人外周血调节性T细胞(Treg)、Th1细胞、Th2细胞、调节性B细胞(Breg)、树突状细胞(DC)等免疫细胞亚群,并进行比较。结果与正常对照组相比,GD患者Treg比例降低,Th1细胞、Th2细胞数量增加;Breg数量减少;DC数量下降,髓样DC(mDC)占DC比例下降,而浆细胞样DC(pDC)比例升高,成熟型DC比例升高。结论 GD患者外周血中存在免疫调节细胞的失衡,可能在GD的发生、发展中起一定作用。     

鼻息肉患者外周血Th17/Treg细胞比率的失衡及其临床意义

目的:观察鼻息肉(NP)患者外周血Th17和Foxp3+ CD4+ CD25+调节性T细胞(Treg)的水平及其与临床病情的关系,初步探讨Th17/Treg细胞的比率失衡在NP发病机制中的作用和意义.方法:NP患者根据鼻内镜和鼻窦CT评分的结果分为1组(鼻内镜检查评分:2～8分;CT检查评分:3～10分,n=23)和2组(鼻内镜检查评分:8～12分;CT检查评分:10～19分,n=23),选取10例单纯鼻中隔偏曲患者作为对照组.采用流式细胞术检测外周血中Th17和Treg的比例,并对Th17/Treg的比率及鼻内镜、鼻窦CT评分进行相关性分析.结果:两组NP患者外周血中Th17细胞比率明显高于对照组(P＜0.01),且2组高于1组(P＜0.05).两组NP患者外周血中Treg细胞的比率明显低于对照组(P＜0.01),但两组间差异无统计学意义.两组NP患者Th17/Treg细胞的比率明显高于对照组(P＜0.01),且两组间差异显著(P＜0.05).同时,相关性分析结果显示,Th17/Treg细胞的比率与鼻内镜、鼻窦CT评分呈正相关(r=0.562,r=0.667,P＜0.01).结论:NP患者外周血中Th17细胞比率增加和Treg细胞比率降低所致的Th17/Treg细胞比率失衡,可能在NP的发生发展中起着重要作用.Th17/Treg比率失衡的程度可能与临床病情严重程度密切相关.     

免疫抑制剂对Graves病患者免疫调节细胞的体内外作用研究

目的观察Graves病(Graves disease,GD)患者外周血免疫调节细胞在体外对地塞米松(DEX)、硫唑嘌呤(AZA)及环磷酰胺(CTX)等免疫抑制剂的反应及其体内在甲状腺局部注射DEX后的变化,探讨DEX等免疫抑制剂在甲状腺局部注射治疗GD过程中对免疫调节系统的作用。方法 1)用DEX、AZA及CTX等免疫抑制剂处理GD患者外周血单个核细胞,采用流式细胞术检测调节性T细胞(Treg)、辅助性T细胞(Th)、调节性B细胞(Breg)、树突状细胞(DC)等免疫细胞亚群变化。2)21例抗甲状腺药物治疗维持期的GD患者,甲状腺局部注射DEX,治疗前后分别检测外周血Treg、Th、Breg、DC等免疫细胞亚群变化。结果 1)免疫抑制剂处理后,各组Treg水平均减少(P0.05),DEX组Th2细胞减少(P0.05),CTX组Th1细胞增加(P0.05);DEX组Breg增多(P0.05),其余2组无明显统计学差异;各组DC细胞中,mDC均明显升高伴pDC下降,DEX、CTX组(P0.05),AZA组(P0.01),DEX组成熟型CD83+及HLA-DR+DC均下降(P0.05),其余2组无明显统计学差异。2)甲状腺局部注射DEX后,与注射前相比,GD患者外周血中Treg细胞增多(P0.05),Th2细胞减少(P0.05),Th1细胞无明显变化(P0.05);Breg比例升高(P0.05);DC亚群中,mDC升高伴pDC下降(P0.05),成熟型CD83+及HLA-DR+DC比例均下降(P0.05)。结论 DEX能有效调节GD患者外周血Treg、Breg及DC等调节性细胞及其亚群水平,重建Th1/Th2平衡,可作为甲状腺局部注射免疫抑制剂治疗GD的首选用药。     

子痫前期患者外周血树突状细胞与T细胞亚群的变化

目的 观察子痫前期患者外周血中的树突状细胞亚群与T细胞亚群相关细胞因子的变化.方法 实验组为子痫前期患者32例,对照组为未孕妇女20例,正常妊娠妇女20例.采集研究对象外周血细胞,流式细胞术检测全血细胞中髓系树突状细胞(mDC)和浆细胞样树突状细胞(pDC);分离外周血单个核细胞,经胞内细胞染色检测Th1、Th2、Th17细胞数量及Th1/Th2比值.结果 子痫前期组mDC百分比(0.33±0.12)％和mDC/pDC比值(2.96±1.65)均高于正常妊娠组,二组数据有明显差异(P＜0.05);pDC百分比(0.16±0.13)％较正常妊娠组(0.21 ±0.12)％有所下降,二组差异有统计学意义(P＜0.05).子痫前期组IFN-γ、IL-4和IL-17的百分比分别为(18.67 ±1.96)％、(1.88±0.51)％和(1.36±0.59)％,与正常妊娠组相比均有显著性差异(P＜0.01).子痫前期组mDC/pDC比率和Th1/Th2之间呈显著正相关(r=0.637,P＜0.01);Th17表达率与pDC表达率之间呈负相关(r=-0.670,P＜0.05),与mDC/pDC比率之间呈显著正相关(r=0.772,P＜0.01).结论 子痫前期患者外周血中树突状细胞亚群和Th1、Th2、Th17型细胞因子异常表达,可能是患者发生免疫失衡的重要原因.     

Th3、Th17及相关细胞因子在自身免疫性甲状腺疾病发病中的作用

目的:研究Th17、Th3细胞及相关细胞因子IL-17、TGF-pI在自身免疫性甲状腺疾病(AITD)发病机制中的作用及意义.方法:收集49例初诊的自身免疫性甲状腺疾病患者,分为Graves’病(GD)(n=30)组和桥本甲状腺炎(HT)(n=19)组,另选年龄、性别匹配的18个正常人作为健康对照.Graves’病组采用甲巯咪唑治疗,桥本甲状腺炎组采用甲状腺素钠治疗,均随访至甲状腺功能正常.采用流式细胞仪技术检测外周血中Th17和Th3细胞比例.采用ELISA方法检测血浆IL-17、TGF-β1水平.结果:GD和HT初诊组患者外周血Th17细胞较正常对照组明显升高(P＜0.05).Th3细胞在HT初诊组中明显升高,而其在GD初诊组中明显下降(P＜0.05).血浆IL-17水平在GD和HT初诊组、缓解组均较正常对照组明显升高(P＜0.05).血浆TGF-β1水平在HT初诊组和缓解组均明显升高(P＜0.05).结论:Th17、Th3细胞和相关细胞因子IL-17、TGF-β1可能参与了自身免疫性甲状腺疾病的发病,TGF-β1可能在HT的发病中起了一个促炎因子的作用.     

Graves'病和桥本氏甲减患者外周血Th1、Th2、Th17、Treg细胞的变化

目的:探讨Graves'病和桥本氏甲减患者外周血中Th1、Th2、Th17细胞和CD4+CD25+FOXP3+调节性T细胞(Treg)的比例及其意义.方法:选择新诊断但未治疗的Graves'病(GD)患者25例、桥本氏甲减25例和正常对照者25例,通过流式细胞技术测定其外周血中Th1、Th2、Th17和CD4+CD25...     

肺癌患者外周血T淋巴细胞亚群变化特点及临床意义

目的:探讨肺癌患者外周血T淋巴细胞变化及其临床意义。方法:采用流式细胞术检测法检测56例肺癌患者(其中20例早中期肺癌、36例晚期肺癌);18例健康人群外周血单个核细胞(Peripheral blood mononuclear cells,PBMC)中总T细胞(CD3+)、Th细胞(CD3+CD4+)、Tc细胞(CD3+CD8+)、NKT细胞(CD3+CD16+CD56+)、NK细胞(CD3-CD16+CD56+)、调节性T细胞(Treg,CD4+CD25+Foxp3+)占CD4+T细胞比例和CD3+γδT细胞比例。结果:56例肺癌患者总T细胞(CD3+)比例明显低于健康组(61.41%±7.88%vs71.63%±5.59%,P0.001),肺癌患者Tc细胞比例明显低于健康组(23.58%±7.18%vs28.44%±5.20%,P0.05),肺癌患者Treg比例明显高于健康组(6.20%±1.63%vs3.65%±2.00%,P0.001);肿瘤组外周血CD3+γδT细胞比例显著低于健康组(3.35%±1.41%vs5.53%±1.87%,P0.01)。但肺癌患者晚期组与肺癌早中期组比较外周血CD3+γδT细胞比例显著增高(3.70%±1.89%vs2.64%±1.41%,P0.05),肺癌患者晚期组与早中期组比较外周血Treg细胞比较显著增高(6.78%±2.64%vs5.06%±1.22%,P0.05)。肺癌患者外周血中NK细胞低于健康组(15.02%±7.61%vs18.74%±6.39%,P0.05),而Th细胞和NKT细胞都有所降低,但差异尚不具统计学意义(P0.05)。结论:肺癌患者总T细胞、Th细胞、Tc细胞和NK细胞都有所降低,Treg细胞有所上升,提示肺癌患者处于免疫抑制状态。Treg细胞和CD3+γδT细胞与肺癌的临床病程具有一定的关联性。     

慢性淋巴细胞性甲状腺炎患者外周血Th17的检测及意义

目的:探讨慢性淋巴细胞性甲状腺炎(CLT)患者外周血辅助性T细胞17(Th17)的水平及意义.方法:收集甲状腺功能正常组CLT患者15例,甲状腺功能减退组CLT患者30例及20例健康对照组研究者的外周血样.采用胞内细胞因子染色流式细胞术(FCM)分析法,检测各组外周血Th17细胞占CD4+T细胞比值.应用电化学发光法检测CLT患者及健康对照组血清中抗甲状腺过氧化物酶抗体(TPO-Ab)和抗甲状球蛋白抗体(TG-Ab)水平.结果:与健康对照组比较:甲状腺功能正常组及甲状腺功能减退组的CLT患者外周血Th17/CD4+T细胞比值分别为(3.64±1.96)%、(3.13±1.56)%,均高于健康对照组(0.50±0.32%,P<0.01);两组CLT患者血清TPO-Ab和TG-Ab水平分别为(398.99±222.88;1456.02±1054.97)IU/L和(423.26±167.21;1587.94±1210.36)IU/L,均明显高于健康对照组(15.23±10.36;36.32±26.99)IU/L;差异有统计学意义(P<0.01).CLT患者外周血Th17/CD4+T细胞比值改变与血清TPO-Ab及TG-Ab阳性表达呈显著正相关(分别为r=0.50,r=0.43;P<0.01).结论:两组CLT患者外周血Th17细胞比率增加,且与甲状腺自身抗体水平存在显著正相关,Th17细胞可能参与CLT的发生发展.     

强直性脊柱炎患者Th17细胞与CD4+CD25+FoxP3+调节性T细胞的变化及意义

目的 探讨强直性脊柱炎( ankylosing spondylitis,AS)患者Th17和CD4+ CD25+ FoxP3+调节性T细胞比例及相关细胞因子水平的变化及意义.方法 强直性脊柱炎患者40例,正常同年龄对照37例.采用流式细胞术检测外周血Th17与调节性T细胞的比例,双抗体夹心酶联免疫吸附法( ELISA)检测血清IL-6、IL-23、IL-17和TGF-β水平.结果 AS组患者外周血Th17细胞比例明显高于对照组[(1.02±0.34)％vs(0.68:±0.29)％,P＜0.05],CD+ CD25+ FoxP3+细胞比例明显低于对照组[(3.77±0.81)％ vs (4.69±1.23)％,P＜0.05].AS患者血清中IL-6、IL-23、IL-17水平明显高于对照组[ (6.15±2.71) ng/L vs(3.31±1.65) ng/L; (9.44±3.12) ng/ml vs (5.82±2.61) ng/ml;( 10.53±4.97) ng/L vs (6.78±3、26) ng/L,P均＜0.01];差异有统计学意义.与对照组相比,AS组TGF-β水平有下降的趋势[(4,76±2.15) ng/ml vs(5.16±2.02) ng/ml,P＞0.05],但差异无统计学意义.AS患者血清各细胞因子含量与临床及实验室指标无相关性.结论 强直性脊柱炎患者Th17与CD4+CD35+FoxP3+调节性T细胞比例失衡,血清IL-6、IL-23、IL-17和TGF-β水平变化,这些原因可能参与强直性脊柱炎免疫发病过程.     

Graves病患者~(131)I、甲巯咪唑治疗前后外周血Th17细胞及IL-17水平的变化

目的:通过比较研究GD患者131I、甲巯咪唑治疗外周血中Th17细胞、IL-17的变化,探讨Th17细胞在GD发病机制中的作用及意义。方法:收集初诊的GD患者31例采用131I治疗,30例采用甲巯咪唑(MMI)治疗,分别在治疗前(T0)及治疗后1个月(T1)、3个月(T3)进行相关指标测定。另选年龄、性别匹配的29例正常人作为正常对照组。应用流式细胞仪技术检测外周血中Th17细胞比例。采用ELISA法检测各组血清IL-17水平。结果:2种治疗方案T0组外周血Th17细胞及IL-17水平明显高于正常对照组(P0.01)。131I治疗组外周血Th17细胞及IL-17水平在T0、T1、T3组中水平逐渐下降,仍高于正常对照组(P0.01),MMI治疗组变化无统计学差异(P0.05)。采用双变量相关性分析得出Th17细胞比例与IL-17水平呈正相关(r=0.758,P0.05),且两者与甲状腺相关抗体呈正相关。结论:Th17细胞、IL-17在初发GD中高表达,Th17细胞可能参与了GD的发病过程。另外,放射性131I可能通过影响Th17细胞及IL-17起到治疗GD的作用。     

CD52 is a novel costimulatory molecule for induction of CD4+ regulatory T cells

We previously reported that 4C8 monoclonal antibody (mAb) provides a costimulatory signal to human CD4+ T cells and consequently induces regulatory T (Treg) cells, which are hypo-responsive and suppress the polyclonal response of bystander CD4+ cells in a contact-dependent manner. In this study, we identified the antigen of 4C8 mAb as CD52. Costimulation with Campath-1H, a humanized anti-CD52 mAb, also induced Treg cells. Anti-CD52-induced Treg cells suppressed the proliferation of both CD4+ and CD8+ T cells provided with polyclonal or allogeneic stimulation. When Treg cells were induced from Staphylococcal enterotoxin B (SEB) treated cells, they suppressed the response to SEB more efficiently than that to another superantigen, SEA. Furthermore, anti-CD52-induced Treg cells could be expanded by culture with IL-2 followed by CD52-costimulation, and co-injection of expanded Treg cells suppressed lethal xenogeneic graft versus host disease (GvHD) reactions in SCID mice caused by human peripheral blood mononuclear cells (PBMCs).     

Special regulatory T-cell review: Suppressors regulated but unsuppressed

The rise-and-fall and reincarnation of suppressor T cells is reviewed from the perspective of a participant in the field.     

Induction of contact-dependent CD8(+) regulatory T cells through stimulation with staphylococcal and streptococcal superantigens

The bacterial superantigen exotoxins of Staphylococcus aureus and Streptococcus pyogenes are potent stimulators of polyclonal T-cell proliferation. They are the causes of toxic shock syndrome but also induce CD25⁺ FOXP3⁺ regulatory cells in the CD4 compartment. Several studies have recently described different forms of antigen-induced regulatory CD8⁺ T cells in the context of inflammatory diseases and chronic viral infections. In this paper we show that bacterial superantigens are potent inducers of human regulatory CD8⁺ T cells. We used four prototypic superantigens of S. aureus (toxic shock syndrome toxin-1 and staphylococcal enterotoxin A) and Str. pyogenes (streptococcal pyrogenic exotoxins A and K/L). At concentrations below 1 ng/ml each toxin triggers concentration-dependent T-cell receptor Vβ-specific expression of CD25 and FOXP3 on CD8⁺ T cells. This effect is independent of CD4⁺ T-cell help but requires antigen-presenting cells for maximum effect. The cells also express the activation/regulatory markers cytotoxic T-lymphocyte antigen-4 and glucocorticoid-induced tumour necrosis factor receptor-related protein and skin homing adhesins CD103 and cutaneous lymphocyte-associated antigen. Superantigen-induced CD25⁺ FOXP3⁺ CD8⁺ T cells were as potent as freshly prepared naturally occurring CD4⁺ regulatory T cells in suppressing proliferation of CD4⁺ CD25⁻ T cells in response to anti-CD3 stimulation. Although superantigen-induced CD8⁺ CD25⁺ FOXP3⁺ express interleukin-10 and interferon-γ their suppressive function is cell contact dependent. Our findings indicate that regulatory CD8⁺ T cells may be a feature of acute bacterial infections contributing to immune evasion by the microbe and disease pathogenesis. The presence and magnitude of regulatory CD8⁺ T-cell responses may represent a novel biomarker in such infections. Superantigen-induced regulatory CD8⁺ T cells also have therapeutic potential.     

CD8+ suppressor-mediated regulation of human CD4+ T cell responses to glutamic acid decarboxylase 65

Although potentially autoreactive T cells are present even in healthy subjects, most individuals do not develop autoimmune disease. It has been well demonstrated that CD4+ CD25+ regulatory T cells play a significant role in controlling the expansion of autoreactive T cells in the periphery. However, some healthy individuals exhibit measurable responses to self peptide even in the presence of CD4+ CD25+ regulatory cells. This article describes the regulation of human CD4+ T cell responses to glutamic acid decarboxylase 65 (GAD65), an autoantigen implicated in type-1 diabetes, by autologous CD8+ suppressor T cells. In cells cultured from healthy individuals, the inclusion of autologous CD8+ T cells at physiological levels resulted in a dramatic decrease in the magnitude of in vitro CD4+ T cell responses to GAD65 peptide. Based on transwell experiments, the observed suppression was cell contact-dependent. However, antibody blocking studies indicated that suppression was mediated by IL-10. Cell fractionation studies suggested that CD8+ suppressor T cells originate from the CD45RA+ CD27- population. The suppression of CD4+ T cell responses to GAD65 in healthy individuals raises the possibility that CD8+ suppressor T cells play an important role in controlling potentially autoreactive T cells in the general population.     

The Tregs' world according to GARP

Naturally occurring CD4⁺CD25^high regulatory T cells (nTreg) are essential for maintaining tolerance. FOXP3 has been established as a molecular marker of nTreg; however, FOXP3 cannot be used as a reliable marker for bona fide human nTreg since effector T cells also up‐regulate FOXP3 expression upon activation. Despite the important function of nTreg, the underlying molecular mechanisms of nTreg‐mediated suppression are far from defined. Previous studies have demonstrated that the TGF‐β latency‐associated peptide (LAP) is expressed on the surface of nTreg, and that immunosuppression can be mediated by membrane TGF‐β; however, it remains unknown how LAP is bound to nTreg and what is the functional significance of its selective expression on activated nTreg. The nTreg's world may now change according to GARP, an orphan toll‐like receptor composed of leucine‐rich repeats. In this issue of the European Journal of Immunology, a study provides further demonstration that GARP is selectively expressed only in activated human nTreg and nTreg cell clones but not in activated effector T cells, confirming GARP as a bona fide nTreg marker. In addition, GARP binds directly to LAP; yet, GARP over‐expression is insufficient to induce modification of latent TGF‐β into active TGF‐β further clarifying its role in nTreg‐mediated suppression.     

Activated T lymphocytes in pre-eclampsia

PROBLEM: The aim of our study was to investigate the activation markets of T CD3(+), T helper CD4(+) and T cytotoxic CD8(+) cells, as well as, the populations of T na?ve CD4(+) CD45RA(+), T memory CD4(+) CD45RO(+) and T regulatory lymphocytes in PE and healthy pregnant women. METHOD OF STUDY: Twenty-five patients with PE and thirty healthy third trimester pregnant women were included in the study. Peripheral blood mononuclear cells were isolated from peripheral blood, stained with monoclonal antibodies and estimated using the flow cytometric method. RESULTS: The percentages of CD4(+)CD25(+), CD4(+)CD25(dim), CD3(+)HLA-DR(+), CD4(+)HLA-DR(+) and CD8(+)HLA-DR(+) cells did not differ between study groups. The population of T regulatory CD4(+)CD25(bright) lymphocytes was significantly lower in the group of patients with PE when compared with the controls (P < 0.01). The percentages of CD3(+)CD25(+) (P < 0.05), CD8(+)CD25(+) (P < 0.05), CD4(+)45RO(+) (P < 0.01) lymphocytes were significantly higher, while CD4(+)CD45RA(+) (P < 0.01) cells--significantly lower in peripheral blood of patients with PE when compared with the control group. CONCLUSION: The increased levels of T CD4(+)45RO(+) and T CD8(+) CD25(+) cells can suggest the activation of CD4(+) and CD8(+) T lymphocytes in pre-eclampsia. It seems possible that the activation of T lymphocytes is associated with the deficiency of T regulatory cells in PE.     

Natural killer T-cell characterization through gene expression profiling: an account of versatility bridging T helper type 1 (Th1), Th2 and Th17 immune responses

Natural killer T (NKT) cells constitute a distinct lymphocyte lineage at the interface between innate and adaptive immunity, yet their role in the immune response remains elusive. Whilst NKT cells share features with other conventional T lymphocytes, they are unique in their rapid, concomitant production of T helper type 1 (Th1) and Th2 cytokines upon T-cell receptor (TCR) ligation. In order to characterize the gene expression of NKT cells, we performed comparative microarray analyses of murine resting NKT cells, natural killer (NK) cells and naïve conventional CD4⁺ T helper (Th) and regulatory T cells (Treg). We then compared the gene expression profiles of resting and alpha-galactosylceramide (αGalCer)-activated NKT cells to elucidate the gene expression signature upon activation. We describe here profound differences in gene expression among the various cell types and the identification of a unique NKT cell gene expression profile. In addition to known NKT cell-specific markers, many genes were expressed in NKT cells that had not been attributed to this population before. NKT cells share features not only with Th1 and Th2 cells but also with Th17 cells. Our data provide new insights into the functional competence of NKT cells which will facilitate a better understanding of their versatile role during immune responses.     

CD28 costimulation: Walking the immunological tightrope

CD28 function has typically been associated with the generation of effector T‐cell responses to Ag. However, it is also clear that CD28 plays an important role in Treg‐cell biology. Understanding which functions predominate is important when designing therapeutic interventions based on CD28 targeting. An article by Hünig and colleagues [Eur. J. Immunol. 2013. 43: 188–193] in this issue of the European Journal of Immunology uses an inducible gene deletion approach to reveal that, in the steady state, Treg cells intrinsically require CD28 signals for their maintenance in the periphery, whereas homeostasis of conventional T cells is relatively unaffected. Here we highlight the delicate balance created by the ability of CD28 to modulate both regulatory and effector T‐cell responses.